| Background:Injectable autologous fat tissue transplantion of liposuction is a common method in softtissue reconstruction, but it often needs repeat injections or over-filling due tounpredictable absorption rate (20%～90%). In earlier studies, an improve of survivalpercentage when PRF and (or) SVF was added to adipose granule before transplantion invitro studies and vivo studies in rabbits.Purpose:Establishedment of fat transplantation model in minipig, preparation of injectiveadipose granule, PRF and SVF composites, subcutaneous injection of the composites todifferent sites of the animal model. The absorption rates of the experimental group and thecontrol group were calculated and compared to evaluate this news ways of adipose tissuetransplantion.Methods:1. Establishedment of fat transplantation model in minipig: a. the preparation of injective adipose granule, PRF and SVF composites; b. subcutaneous injection of thecomposites in the ears of minipigs; c. injection of the composites in the cavity in theabdominal prepared by a soft tissue expander; d. subcutaneous injection of the compositesin the soft tissue defect in the back of the minipigs.2. Caliper and B-mode ultrasound were used to measure the volume changes of thegraft materials.3. The tissue specimens were prepared for histological examination: hematoxylin andeosin (H&E) staining; Oil-red O staining; Factor VIII immunohistochemical staining forVessel count; Cell apoptosis using Tunel technique. Differences between the experimentalgroup and the control group were analyzed.Results:1. The fat transplantation models in minipig were established: a. the preparation ofinjective fat particles, PRF and SVF composites; b. subcutaneous injection of thecomposites in the ears of minipigs; c. injection of the composites in the cavity in theabdominal prepared by a soft tissue expander; d. subcutaneous injection of the compositesin the soft tissue defect in the back of the minipigs.2. Subcutaneous injection of the composites in the ear of minipigs: The ratio of controlgroup, experimental group A and group B was1︰1︰1, each group included6cases.When20ml of the adipose composites was injected, the diameter of the donor site wasabout5cm, the height of the donor site was about2.5cm, the survival percentage of thecontrol group, experiment group A and experiment group B was6.25%(SD3.59),9.27%(SD3.69) and12.96%(SD4.02), respectively; When10ml of the adipose composites wasinjected, the diameter of the donor site was about5cm, the height of the donor site wasabout1.3cm, the survival percentage of the control group, experiment group A andexperiment group B was10.28%(SD2.42),21.88%(SD4.34) and31.00%(SD4.29),respectively; When5ml of the adipose composites was injected, the diameter of the donorsite was about5cm, the height of the donor site was about0.9cm, the survival percentageof the control group, experiment group A and experiment group B was31.01%(SD10.76),30.80%(SD7.01) and46.75%(SD9.04), respectively.Single dependent variable multi-factor ANOVA analyses show that the surviralpercentage of experiment group B was higher than experiment group A, the surviralpercentage of experiment group A was higher than control group; the surviral percentage of5ml group was higher than10ml group, the surviral percentage of10ml group washigher than20ml. All the differences between groups were statistically significant.The vessel counts of the specimen after Factor VIII immunohistochemical stainingwere anaylized by Single dependent variable multi-factor ANOVA, the results show that in20ml graft groups, the experiment group B had higher vessel count than the experimentgroup A, the experiment group A had higher vessel count than the control group, while in10ml and5ml graft groups, the control group count highest, then the experiment group A,the experiment group B count lowest. The differences between groups were statisticallysignificant.Cell apoptosis using Tunel technique showed that there were a lot more apoptotic cellsin control groups than experimental groups, but no obvious difference between5ml,10mland20ml groups.Oil-red O staining of the specimen showed that the fat lipid droplets were dyed red,the fat bubbles consistency was poor in control groups, the fat bubbles consistency wasbetter in experiment groups, but no siginificant difference between the experiment group Aand group B.3. Injection of the composites in the cavity in the abdominal prepared by soft tissueexpander:9models were set up,3for control group,3for experiment group A, and3forexperiment group B, but all the cases were found severe absorbed, no obvious adiposetissue were found during ultrasound exam and sample surgery, no statistics differencebetween experiment group A and group B.4. Subcutaneous injection of the composites in the soft tissue defect in the back of theminipigs:18cases of the models were set up,6for control group,6for experiment groupA, and6for experiment group B. In the control group, the volume of tissue graft was13.60ml (SD3.05), the thickness of the adipose tissue before the transplantation was13.85mm (SD2.71), and the augmented soft tissue was2.85mm (SD0.82); In theexperiment group A, the volume of tissue graft was11.00ml (SD2.24), the thickness ofthe adipose tissue before the transplantation was12.74mm (SD1.69), and the augmentedsoft tissue was11.27mm (SD5.51); In the experiment group B, the volume of tissue graftwas11.20ml (SD1.30), the thickness of the adipose tissue before the transplantation was13.35mm (SD2.00), and the augmented soft tissue was16.73mm (SD3.27).There was no statistic difference between the groups of volume of tissue graft, andthere was no statistic difference between the groups of the thickness of the adipose tissue before the transplantation, but there was statistic difference between the groups of theaugmented soft tissue.The survival percentage of the augmented thickness of adipose tissue of the controlgroup, the experiment group A and B was11.04%(SD2.80),40.94%(SD16.79) and56.80%(10.19) respectively. ANOVA analyse showed there was statistic differencebetween groups.The vessel count showed that the experiment group A and B had less vessel numberthan control group. The difference between the experiment group A and B was that the cellnumber of each vessel count less in the experiment group B than group A, which maightindicate the adipose tissue in experiment group B was more mature and stable.Conclusions:1. Using minipig as one of the subjects of adipose tissue transplant is a viable option.2. Subcutaneous injection of5ml,10ml and20ml of the composites in the ears ofminipigs shows injection of adipose granule+PRF and adipose granule+PRF+SVF havebetter result than injection of adipose granule only, and injection of adiposegranule+PRF+SVF shows the best results.3. The idea of injection of the composites in the cavity in the abdominal prepared bya soft tissue expander may not be a viable option.4. Experiments of subcutaneous injection of the composites in the soft tissue defect inthe back of the minipigs show that adipose granule+PRF+SVF composites have betterresult in adipose transplantation.5. When PRF and (or) SVF is added in adipose granule during transplantation, thesurvival percentage of the graft material is greatly increased, and the augmented tissuelooks more autologous tissue alike, maybe more mature and stable. |
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